The role of PKCepsilon in diabetic retinopathy

PKCepsilon 在糖尿病视网膜病变中的作用

• 批准号: 8018061
• 负责人: Christian Rask-Madsen
• 金额: $ 12.79万
• 依托单位: JOSLIN DIABETES CENTER
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-02-01 至 2013-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8018061
• 关键词: 5' -AMP-activated protein kinase; Acute; Animal Model; Biology; Cell Culture Techniques; Cell Proliferation; Cell physiology; Cells; Cellular biology; Complications of Diabetes Mellitus; Data; Development; Diabetes Mellitus; Diabetic Retinopathy; Disease; Down-Regulation; Drug Delivery Systems; Endothelial Cells; Endothelium; Evaluation; Eye; Fellowship; Genetic; Growth Factor; Injection of therapeutic agent; Institutes; K-Series Research Career Programs; Knock-out; Knockout Mice; Knowledge; Laboratories; Mediating; Mediator of activation protein; Mentors; Minor; Modeling; Molecular; Nitric Oxide; Outcome; Pathway interactions; Patients; Physicians; Play; Postdoctoral Fellow; Prevention; Process; Production; Protein Isoforms; Protein Kinase C; Proteins; Rattus; Research; Research Personnel; Retina; Retinal; Retinal Diseases; Retinal Neovascularization; Role; Scientist; Signal Transduction; Signaling Molecule; Small Interfering RNA; Streptozocin; Testing; Training; Transgenic Mice; Up-Regulation; Vascular Endothelial Growth Factor Receptor; Vascular Endothelial Growth Factor Receptor-2; Vascular Endothelial Growth Factors; Vascular Permeabilities; Wild Type Mouse; Work; angiogenesis; diabetic; diabetic rat; human NOS3 protein; inhibitor/antagonist; innovation; macular edema; mouse model; neovascularization; novel; overexpression; prevent; programs; proliferative diabetic retinopathy; receptor; research study

DESCRIPTION (provided by applicant): Project summary. The applicant is a physician-scientist who has completed his post-doctoral fellowship in the laboratory of Dr. George L. King, who will continue to function as a mentor while the applicant expands his training in new fields, including retinal endothelial cell biology and animal models of retinal complications of diabetes. This training will be co-mentored by Dr. Lloyd Paul Aiello at the Beetham Eye Institute at Joslin Diabetes Center. The Career Development Award would allow the applicant to become an independent researcher in the field of retinal disease and help achieve the long-term objective of understanding growth factor actions in diabetic eye complications. In diabetic macular edema and proliferative diabetic retinopathy, upregulation of vascular endothelial growth factor (VEGF) plays a central role. Protein kinase C (PKC) isoforms a, (3, and e are activated in the diabetic retina, but so far there has been a lack of data from animal models with isoform-specific manipulation of PKC function. In preliminary results, downregulation of PKCe with small interfering RNA (siRNA) in endothelial cell culture had a dramatic effect on several pathways of VEGF-stimulated signaling and cell function, including nitric oxide production and cell proliferation, with minor or opposite effects seen after downregulation of PKCa, (3, or 8. Further, intravitreal injection of PKCe siRNA inhibited VEGF-stimulated retinal vascular permeability in rats. Therefore, the central hypothesis of this application is that PKCe isoform has a quantitatively major role in mediating VEGF-stimulated Akt, eNOS, and AMP-activated protein kinase (AMPK) signaling, and is critical for development of macular edema and proliferative retinopathy in diabetes. The specific aims are: i. To characterize mechanisms of activation of PKCe in retinal endothelial cells and cell function regulated by PKCe; 2. To determine the effect of PKCe knockout or overexpression on vascular permeability during VEGF stimulation and in diabetes; 3. the effect of knockout or overexpression of PKCe function on ischemic neovascularization in the retina. Relevance. The proposed research is expected to have a positive impact on the prevention of macular edema and proliferative retinopathy in patients with diabetes because it will establish the usefulness of the signaling molecule PKCe as a key mediator of these processes and a potential drug target for these conditions.

描述（由申请人提供）：项目摘要。申请人是一位医师科学家，已在 George L. King 博士的实验室完成了博士后研究金，在申请人扩展其在新领域（包括视网膜内皮细胞生物学和糖尿病视网膜并发症的动物模型。该培训将由乔斯林糖尿病中心 Beetham 眼科研究所的 Lloyd Paul Aiello 博士共同指导。职业发展奖将使申请人成为视网膜疾病领域的独立研究人员，并帮助实现了解生长因子在糖尿病眼部并发症中的作用的长期目标。在糖尿病性黄斑水肿和增殖性糖尿病视网膜病变中，血管内皮生长因子（VEGF）的上调发挥着核心作用。蛋白激酶 C (PKC) 异构体 a、(3 和 e) 在糖尿病视网膜中被激活，但迄今为止，还缺乏对 PKC 功能进行异构体特异性操作的动物模型数据。初步结果显示，PKCe 的下调内皮细胞培养物中的小干扰 RNA (siRNA) 对 VEGF 刺激的信号传导和细胞功能的多个途径产生了巨大影响，包括一氧化氮的产生和细胞增殖，而在PKCa 下调，（3 或 8）。此外，玻璃体内注射 PKCe siRNA 抑制大鼠 VEGF 刺激的视网膜血管通透性。因此，本申请的中心假设是 PKCe 亚型在介导 VEGF 刺激的 Akt 中具有定量的主要作用、eNOS 和 AMP 激活蛋白激酶 (AMPK) 信号传导，对于糖尿病黄斑水肿和增殖性视网膜病变的发展至关重要。具体目标是： i. 表征视网膜内皮细胞中 PKCe 的激活机制以及 PKCe 调节的细胞功能； 2. 确定 PKCe 敲除或过表达对 VEGF 刺激期间和糖尿病中血管通透性的影响； 3. PKCe功能敲除或过表达对视网膜缺血性新生血管的影响。关联。拟议的研究预计将对糖尿病患者黄斑水肿和增殖性视网膜病变的预防产生积极影响，因为它将确立信号分子 PKCe 作为这些过程的关键介质和这些疾病的潜在药物靶点的有用性。